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Signal Transduction Protein Phosphorylation Ser / Thr Kinases Other Kinases

Human MLKL ELISA Kit (ab263892)

Human MLKL ELISA Kit (ab263892)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 6.29 pg/ml
  • Range: 78.13 pg/ml - 5000 pg/ml
  • Sample type: Cell Lysate
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Overview

  • Product name

    Human MLKL ELISA Kit
    See all MLKL kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Extract 8 10.2%
    Inter-assay
    Sample n Mean SD CV%
    Extract 3 6.8%
  • Sample type

    Cell Lysate
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    6.29 pg/ml
  • Range

    78.13 pg/ml - 5000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell Lysate 90 82% - 100%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Human MLKL ELISA Kit (ab263892) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of MLKL protein in cell lysate. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human MLKL with 6.29 pg/ml sensitivity.


    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:


            - Single-wash protocol reduces assay time to 90 minutes or less
            - High sensitivity, specificity and reproducibility from superior antibodies
            - Fully validated in biological samples
            - 96-wells plate breakable into 12 x 8 wells strips


    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

  • Notes

    Mixed lineage kinase domain-like (MLKL) is a pseudokinase containing a protein kinase-like domain that plays a critical role in tumor necrosis factor (TNF)-induced necroptosis as a functional substrate of receptor-interacting protein kinase 3 (RIP3). MLKL, together with receptor-interacting protein 1 (RIP1) and RIP3, form a necrosis signaling complex termed the necrosome, within which MLKL is phosphorylated at T357 and S358 residues by RIP31. Once phosphorylated, MLKL oligomers localize to the plasma membrane and form disrupting pores, capable of regulating the influx of Na+ and Ca2+ rupturing the cell membrane. Necroptosis requires MLKL and RIP3 and is implicated in the pathogenesis of various diseases through its role in necroptosis. Human MLKL shares 69% and 68% sequence homology with mouse and rat, respectively.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human MLKL Capture Antibody 1 x 600µl
    10X Human MLKL Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 5BI 1 x 6ml
    Human MLKL Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Cancer
    • Cell Death
    • Necroptosis
  • Sequence similarities

    Belongs to the protein kinase superfamily.
    Contains 1 protein kinase domain.
  • Domain

    The protein kinase domain is predicted to be catalytically inactive.
  • Target information above from: UniProt accession Q8NB16 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • 9130019I15Rik
    • FLJ34389
    • hMLKL
    • Mixed lineage kinase domain like
    • Mixed lineage kinase domain like protein
    • Mixed lineage kinase domain like pseudokinase
    • Mixed lineage kinase domain-like protein
    • Mlkl
    • MLKL_HUMAN
    see all
  • Database links

    • Entrez Gene: 197259 Human
    • Omim: 615153 Human
    • SwissProt: Q8NB16 Human
    • Unigene: 119878 Human

    Images

    • SimpleStep ELISA Protocol Diagram
      SimpleStep ELISA Protocol Diagram

      SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

    • Example of Human MLKL standard curve in 1X Cell Extraction Buffer PTR.
      Example of Human MLKL standard curve in 1X Cell Extraction Buffer PTR.

      The MLKL standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

    • Interpolated concentrations of native MLKL in Human cell culture extract based on a 75 µg/mL extract load.
      Interpolated concentrations of native MLKL in Human cell culture extract based on a 75 µg/mL extract load.

      The concentrations of MLKL were measured in duplicate and interpolated from the MLKL standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MLKL concentration was determined to be 741.83 pg/mL in HUVEC cell extract.

    • Sandwich ELISA - Human MLKL ELISA Kit (ab263892)
      Sandwich ELISA - Human MLKL ELISA Kit (ab263892)
      To learn more about the advantages of recombinant antibodies see here.

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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